Table 1 Kinetic parameters and coupling free energies (ΔΔΔG) of the binding reaction of C-SH3 Grb2 wild-type and its site-directed mutants with Gab2503–524 wild-type, Gab2503–524 P510A and Gab2503–524 P512A.

From: Mapping the allosteric network within a SH3 domain

C-SH3 Grb2

Gab2503–524 WT

Gab2503–524 P510A

Gab2503–524 P512A

mutant

kon (µM−1 s−1)

koff (s−1)

KD (µM)

kon (µM−1 s−1)

koff (s−1)

KD (µM)

∆∆∆G (kcal mol−1)

Distance (Å)

kon (µM−1 s−1)

koff (s−1)

KD (µM)

∆∆∆G (kcal mol−1)

Distance (Å)

WT

25.0 ± 1.8

40 ± 4

1.7 ± 0.1

27.4 ± 2.2

50 ± 4

1.6 ± 0.1

—

21.4

18.5 ± 1.0

70 ± 7

3.8 ± 0.1

—

23.8

T1S

25.3 ± 1.6

60 ± 6

2.3 ± 0.1

30.6 ± 1.9

50 ± 5

1.5 ± 0.1

0.22 ± 0.08

23.0

22.5 ± 0.8

100 ± 6

4.3 ± 0.1

0.11 ± 0.06

27.4

Y2A

22.9 ± 2.1

50 ± 5

2.0 ± 0.1

22.9 ± 1.3

90 ± 10

3.7 ± 0.1

−0.37 ± 0.07

15.7

16.0 ± 1.0

90 ± 6

5.8 ± 0.1

−0.13 ± 0.07

18.6

V3A

32.2 ± 3.9

30 ± 3

0.9 ± 0.2

26.6 ± 2.9

60 ± 6

2.1 ± 0.2

−0.48 ± 0.12

10.4

27.9 ± 2.5

70 ± 7

2.4 ± 0.1

−0.07 ± 0.11

16.9

A5G

24.0 ± 1.4

60 ± 10

2.3 ± 0.2

30.5 ± 3.4

60 ± 6

1.9 ± 0.2

0.09 ± 0.09

9.9

14.7 ± 2.0

100 ± 14

6.4 ± 0.2

−0.13 ± 0.07

13.1

L6A

33.9 ± 2.4

40 ± 20

1.3 ± 0.4

14.9 ± 5.0

120 ± 30

8.1 ± 0.4

−1.06 ± 0.18

5.1

13.1 ± 2.3

110 ± 16

8.5 ± 0.2

−0.62 ± 0.18

4.9

F7A

—

—

—

—

—

—

—

10.4

—

—

—

—

10.2

F19A

—

—

—

—

—

—

—

18.6

—

—

—

—

24.6

F24A

22.3 ± 1.8

50 ± 5

2.3 ± 0.1

16.7 ± 2.3

60 ± 6

3.7 ± 0.2

−0.28 ± 0.07

13.7

14.7 ± 0.9

90 ± 6

6.0 ± 0.1

−0.07 ± 0.06

18.0

I25V

23.3 ± 1.3

40 ± 10

1.6 ± 0.3

23.4 ± 0.5

60 ± 3

2.7 ± 0.1

−0.32 ± 0.11

25.3

13.7 ± 1.6

90 ± 9

6.4 ± 0.2

−0.32 ± 0.11

28.0

H26A

26.3 ± 3.5

80 ± 8

3.1 ± 0.2

17.7 ± 0.9

80 ± 7

4.3 ± 0.1

−0.21 ± 0.07

25.7

16.9 ± 1.5

80 ± 10

4.4 ± 0.2

0.26 ± 0.07

23.1

S31A

32.5 ± 2.0

20 ± 2

0.7 ± 0.1

15.5 ± 1.2

90 ± 8

5.6 ± 0.1

−1.20 ± 0.11

21.7

23.1 ± 1.3

70 ± 9

3.0 ± 0.1

−0.38 ± 0.11

24.9

A39G

21.5 ± 3.9

70 ± 7

3.4 ± 0.2

28.5 ± 6.4

50 ± 4

1.6 ± 0.2

0.42 ± 0.11

16.4

34.6 ± 4.9

60 ± 6

1.7 ± 0.2

0.86 ± 0.09

23.0

H41A

31.6 ± 3.0

60 ± 5

1.8 ± 0.1

19.8 ± 1.8

50 ± 5

2.3 ± 0.1

−0.17 ± 0.08

23.0

18.0 ± 2.9

80 ± 8

4.3 ± 0.2

−0.04 ± 0.07

24.6

T44S

28.6 ± 1.5

40 ± 5

1.4 ± 0.1

19.8 ± 1.7

50 ± 5

2.5 ± 0.1

−0.32 ± 0.08

4.6

23.2 ± 2.3

70 ± 15

3.0 ± 0.2

0.04 ± 0.09

5.2

Y51A

—

—

—

—

—

—

—

14.2

—

—

—

—

18.0

T53S

34.9 ± 1.8

20 ± 2

0.7 ± 0.1

26.9 ± 1.7

50 ± 5

1.9 ± 0.1

−0.62 ± 0.12

17.7

9.8 ± 1.0

90 ± 7

8.6 ± 0.1

−0.99 ± 0.11

20.7

A54G

24.0 ± 0.7

40 ± 5

1.7 ± 0.1

21.1 ± 1.1

60 ± 8

2.9 ± 0.1

−0.33 ± 0.08

17.6

25.9 ± 4.4

80 ± 8

2.9 ± 0.2

0.15 ± 0.08

21.9

V55A

25.7 ± 1.6

40 ± 4

1.7 ± 0.1

13.8 ± 1.6

100 ± 10

6.9 ± 0.2

−0.81 ± 0.07

21.4

12.3 ± 1.4

80 ± 9

6.4 ± 0.2

−0.29 ± 0.07

23.8

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